Abstract
Two typical medium replacement methods are employed to culture cells, that is, traditional manual medium change, wherein complete medium changes are performed at short-term intervals; and perfusion, wherein continuous culture media delivery and discharge are performed. The former is associated with the issue of readily accumulation of metabolic waste products, which is resolved by the latter. However, the latter requires a specific chamber for cell culturing such as a microfluidic channel for flow stability. Generally, bio-medical researchers require a culturing process that utilizes commercial culture dishes to test their conventional manipulation know-how, experience, and protocols. Thus, we constructed a perfusion-culture system using commercial 35 mm culture dishes. To use commercial culture dishes, it is necessary to maintain the culturing conditions including a constant volume and uniform flow rate, and we invented a novel adapter, culture dish adapter (CD-Adapter), attached to a commercial culture dish. The CD-Adapter is fixed with a holding jig to keep the medium volume constant, so it can maintain a uniform flow in the culture dish. Additionally, we demonstrate the applicability of the system by applying it to a perfusion culture of 293T cells, revealing a 50% reduction in DDIT3, a cellular stress marker, compared with that in conventional manual medium change.
Highlights
Technologies for cultivating animal cells in vitro are called cell culture techniques [1]
We developed a versatile system for perfusion culture that can be equipped with various commercial 35 mm culture dishes
Tube pumps are well suited for perfusion cultures because they are not limited by reservoir size or the amount of time for which they can operate; they have a low minimum flow rate and low dead volume, and do not require any preprocessing of the culture medium
Summary
Technologies for cultivating animal cells in vitro (in an artificial environment) are called cell culture techniques [1]. One of the major advantages of cultivating cells in vitro is that one can control the cell culture environment parameters in the physical environment, such as temperature, pH, and shear stress, as well as those in the physiological environment, including hormone or metabolic waste product concentrations [2,3,4,5,6]. To repeatedly obtain stable experimental results by fully exploiting these advantages, it is necessary to apply methods that effectively culture homogeneous cells with good reproducibility (high-quality cells) [8]. It is necessary to regularly replace the culture medium when cultivating cells. This method of cell culture, known as manual medium change, is widely used [9].
Sign-in/Register to view highlights & summary Sign-in/Register to access full text options 
Talk to us
Join us for a 30 min session where you can share your feedback and ask us any queries you have
Schedule a call
